Agrisera/MIP1 | Aquaporin, glycerol transport activity/AS15 2826/


productinformation
Background		MIP1(Aquaporin,glyceroltransportactivity)islocalizedtothecontractilevacuole,whichinmostfreshwaterflagellatesisusedtoexpelexcesswater.
Immunogen		KLH-conjugatedsyntheticpeptide,derivedfromChlamydomonasreinhardtiiMIP1,UniProt:Q5VLJ9
Host		Rabbit
Clonality		Polyclonal
Clone
Purity		Affinitypurifiedserum
Format		LyophilizedinPBSpH7.4
Quantity		50µg
Reconstitution		Forreconstitutionadd50µlofsterilewater.
Storage		storelyophilized/reconstitutedat-20°C;oncereconstitutedmakealiquotstoavoidrepeatedfreeze-thawcycles.Please,remembertospintubesbrieflypriortoopeningthemtoavoidanylossesthatmightoccurfromlyophilizedmaterialadheringtothecaporsidesofthetubes.
Testedapplications		westernblot(WB),immunogold(IG)
Relatedproducts		Chlamydomonasreinhardtiiantibodycollection Algalproteinextractionbuffer Secondaryantibodies
Additionalinformation

applicationinformation
Recommendeddilution		1:10000withECL(WB),1:200(IG)
Expected\|apparentMW		31.5\|43kDa
Confirmedreactivity		Chlamydomonasreinhardtii(strainsCC3395,andUVM4fromNeupertetal.,2009.(GenerationofChlamydomonasStrainsthatEfficientlyExpressNuclearTransgenes.)
Predictedreactivity		Chlamydomonasreinhardtii
Notreactivein		noconfirmedexceptionsfrompredictedreactivityarecurrentlyknown
Additionalinformation		AsMIP1isamembraneproteinpleaseuseahighredoxpotentialinyourlysisbuffer.
Selectedreferences		Komsic-Buchmannetal.(2014).TheContractileVacuoleasaKeyRegulatorofCellularWaterFlowinChlamydomonasreinhardtii.Eukaryoticcell(13),Issue11:1421-30.

applicationexample

10µgoftotalproteinfrom15mlcellsuspensionofChlamydomonasreinhardtiiCC3395,withacelldensityof(~10times8)extractedwithSDSbuffersupplememntedwithProteaseinhibitor(cOmpleteUltratablets,EDTAfree,Roche,Mannheim),afterremovalofthecelldebrisviacentrifugation,DTTwasadded(100mMfinalconcentration)andwereseparatedon12%SDS-PAGEandblotted1htoPVDFusingtanktransfer.BlotswereblockedwithRotiBlock-Solution(Roth,Darmstadt)for1hatroomtemperature(RT)orat4°CONwithagitation.Blotwasincubatedintheprimaryantibodyatadilutionof1:10,000inTBSwith2%milkpowderfor1hatRTwithagitation.Theantibodysolutionwasdecantedandtheblotwaswashed3timesfor10minuteseachwithTBS-TatRTwithagitation.Blotwasincubatedinsecondaryantibody(anti-rabbitIgGhorseradishperoxidaseconjugated,fromSigma,St.Luis,MO)dilutedto1:80000inTBStith5%milkpowderfor1hatRTwithagitation.Theblotwaswashedasaboveanddevelopedfor1-2minwithECLaccordingtothemanufacturersinstructions.Exposuretimewas40seconds.

CourtesyofDr.KarinKomsic-BuchmannandProf.Dr.BurkhardBecker,UniversityofCologne,Germany

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Agrisera/MIP1 | Aquaporin, glycerol transport activity/AS15 2826/